Environmental and Molecular Mutagenesis - Legacy Tobacco ...

42 1989 EMS Abstracts
http://legacy.library.ucsf.edu/tid/clb93d00/pdf
Notes concurrent increase in mitotic cross-overs was observed, suggesting the disturbance
of chromosome segregation as primary genetic event . Conversely, in the case of hydroquinone
the secondary origin of chromosome malsegregants was demonstrated, whereas
additional assays are required to disclose the nature of mitotic segregants produced
by econazole treatments . Finally, colchicine, diazepam, cadmium chloride and pyrimeta-
mine were assayed in a wide range of doses with completely negative results . (Work
partially granted by the E .E .C . contract n . EV4C-0044-I(a) ) .
116
THE DEVELOPMENT OF A PANEL OF HUMAN CELL LINES EXPRESSING SPECIFIC HUMAN
CYTOCHROME P450 cDNAS
C . L . Crespi and B . W . Penman, Gentest Corporation, Woburn, MA (USA)
We have previously reported a development of human lymphoblastold cell lines which express
tranfected human cytochrome P450IIA3 and microsomal epoxide hydrolase cDNA's . We have
now isolated additional cDNA's encoding human cytochrome P4501IE1, P450IIC9 and P450IIIA4
using oligonucleotide probes . The cDNA's were sized and the DNA sequence of the 5' end of
the isolate was determined. The cDNA's had the following properties :
P450 Form cDNA Size DNA Sequence HomologV Coding Sequence
fIE1 1 .7 kb 97% 11 bp short
11C9 1 .9 kb 100% 32 bp short
IIIA4 2 .1 kb 99% Complete
The IIE1 and IIC9 Isolates which did not contain complete coding sequences were completed
by the addition of synthetic oligonucleotides. The complete cDNA's are being introduced into
a pHEBo based expression vector system under the control of the herpes simplex virus thymidine
kinase gene promoter and polyadenylation signal . Recombinant cell lines expressing these
cytochrome P450 cDNA's are being constructed . These cell lines, in addition to cell lines
expressing human cytochrome P450IIA3 and P4S01A1, should provide a useful panel of cell lines
evaluating the ability of specific human cytochrome P450 forms to activate chemical mutagens .
117
ASSESSMENT OF THE MUTAGENIC EFFECT OF MATERNAL FACTORS ON HUMAN CHORICNIC VILLI BY
MICRONUCLEUS TEST
Y .Q . Cui, Z .W . Dong, S .B . Liu, S .C . Chang, Y . Wang, X .Y . Ji, National Research
Institute for Family Planning, Beijing, People's Republic of China .
Rapid determination of DNA damage by micronucleus test is well accepted . Animal bone
marrow cells or human peripheral lymphocytes used in most studies could not directly
reflect the influence of the mutagenic effect on the offsprings by environmental
factors . Although the cytogenetic effect of human chorionic villi chromaomes are used
in prenatal diagnosis, human chorionic villi sdcronucleus test to detect directly the
mutagenic effect of environmental factors has not been reported in the literature .
Direct determination of human chorionic villi sdcronuclei (CVlIIi) was established by
us, to study the mutagenic effect of mother's age, gravidity, abortion history, contraception,
smoking and drinking on the offsprings . Cross investigation and micronucleus
test were used in 507 couples undergoing artificial abortion . Micronuclei were scored
according to Countryman's standard . 2, 000 interphases were observed in each subject
for CVMN frequency . Aresine transformation {aresine (Sqr(P)1) was used in transforming
CVMN frequency :and the analysis of variance were used for statistics . No correlation
between CVM4 frequency and mother's age, gravidity, gestation age, abortion history,
and contraception was found . Neither smoking nor drinking habit was found among the
women of this study . The CVlA1 frequency of husband smoking was 0 .76+0 .06%., of husband
non-smoking-drinking was 0 .55t0 .06X., of husband drinking was 0 .57t0 .20X ., of husband
smoking and drinking was 0 .79* 0 .08X . . There was a statistical difference in CVMN
frequency between husband smoking and non-saaking . No significant difference was
found between husband drinking and non-drinking .
118
METHYLAZOMETHANOL ACETATE (MAMoAC), A STABLE FORH OF THE ULTIMATE MUTAGEN FROM
CYCASIN IS ACTIVATED BY PORCINE LIVER ESTERASE . Michael L . Cunningham and Timothy F .
McMahon, Systemic Toxicology Branch, National Institutes of Health, NIEHS,
Research Triangle Park, North Carolina, 27709
Nethylazomethanol (HAH) is the short-lived toxic and carcinogenic aglycone of cycasin,
a natural component of the cycad plant . These plants may be consumed by people in the
South Pacific islands either as foodstuff or in medicinal preparations . Thg
spontaneous decomposition of MAN to reactive methyl carbonium ions above 0 C makes
the study of its mutagenic effects difficult . Hovever, we have used the stable ethyl
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